Guasti P N, Monteiro G A, Maziero R R D, Carmo M T, Dell'Aqua J A, Crespilho A M, Rifai E A, Papa F O
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science, São Paulo State University, Botucatu, Brazil.
Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science, São Paulo State University, Botucatu, Brazil.
Anim Reprod Sci. 2017 Apr;179:27-34. doi: 10.1016/j.anireprosci.2017.01.013. Epub 2017 Feb 3.
The aims of this study were to determinate whether pentoxifylline (PTX) increases the motion parameters of fresh and frozen-thawed equine epididymal spermatozoa, to evaluate the tyrosine phosphorylation of frozen-thawed epididymal sperm in the presence of PTX and to determine whether the PTX-treatment of stallion epididymal sperm prior to freezing improves the fertility response of mares to a reduced number of spermatozoa per insemination dose. Fifty epididymis were flushed with a skim milk based extender with or without PTX. The pre-treatment with PTX enhanced the sperm motility after being harvested (P<0.05); however the freeze-thaw process did not alter the sperm kinematics between control and treated samples (P>0.05). Plasma membrane integrity did not differ between control and PTX group after recovery and after thawing (P>0.05), as observed in tyrosine phosphorylation, which the PTX treatment did not alter the percentage of tail-associated immunofluorescence of cryopreserved epididymal sperm (P>0.05). For the fertility trial, different insemination groups were tested: 800×10 epididymal sperm (C800); 100×10 epididymal sperm (C100); 100×10 epididymal sperm recovered in an extender containing PTX (PTX100). The conception rates for C800; C100 and PTX100 were 68.7% (11/16); 31.5% (5/16) and 50% (8/16), respectively. The conception rate did not differ among groups (P>0.05), however, a low number of animals was used in this study. A trend toward significance (P=0.07) was observed between C800 and C100 groups. In conclusion, PTX has no deleterious effect on sperm motility, viability and capacitation of cryopreserved stallion epididymal sperm. The conventional artificial insemination with 100×10 sperm recovered with PTX ensures acceptable conception rates and maximize the limited number of doses of cryopreserved stallion epididymal sperm.
本研究的目的是确定己酮可可碱（PTX）是否能提高新鲜和冻融马附睾精子的运动参数，评估PTX存在时冻融附睾精子的酪氨酸磷酸化情况，并确定在冷冻前对种公马附睾精子进行PTX处理是否能提高母马对每输精剂量减少的精子数量的生育反应。用含或不含PTX的脱脂乳基稀释液冲洗50个附睾。PTX预处理提高了采集后精子的活力（P<0.05）；然而，冻融过程并未改变对照组和处理组样本之间的精子运动学参数（P>0.05）。恢复后和解冻后，对照组和PTX组之间的质膜完整性没有差异（P>0.05），酪氨酸磷酸化情况也是如此，PTX处理并未改变冷冻保存的附睾精子尾部相关免疫荧光的百分比（P>0.05）。对于生育试验，测试了不同的输精组：800×10附睾精子（C800）；100×10附睾精子（C100）；在含PTX的稀释液中回收的100×10附睾精子（PTX100）。C800、C100和PTX100的受孕率分别为68.7%（11/16）、31.5%（5/16）和50%（8/16）。各组之间的受孕率没有差异（P>0.05），然而，本研究中使用的动物数量较少。在C800组和C100组之间观察到有显著差异的趋势（P=0.07）。总之，PTX对冷冻保存的种公马附睾精子的活力、生存能力和获能没有有害影响。用PTX回收的100×10精子进行传统人工授精可确保可接受的受孕率，并最大限度地利用有限数量的冷冻保存种公马附睾精子剂量。
